中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
Chinese Journal of Pathophysiology
2015年
11期
2047-2052
,共6页
陆新华%王辉%宁昕杰%罗骏成%梁家骥
陸新華%王輝%寧昕傑%囉駿成%樑傢驥
륙신화%왕휘%저흔걸%라준성%량가기
周围神经损伤%微小RNA-21%雪旺细胞%细胞凋亡
週圍神經損傷%微小RNA-21%雪旺細胞%細胞凋亡
주위신경손상%미소RNA-21%설왕세포%세포조망
Peripheral nerve injury%MicroRNA-21%Schwann cells%Apoptosis
目的：探讨周围神经损伤时，微小RNA－21（microRNA－21，miR－21）与雪旺细胞（SC）凋亡的关系及相关分子机制。方法：采用实时荧光定量PCR（ real－time PCR）检测动物模型中miR－21以及人第10号染色体缺失的磷酸酶及张力蛋白同源基因（phosphatase and tensin homologue deleted on chromosome 10，PTEN）的表达情况；通过将miR－21类似物（miR－21－mimic）、miR－21抑制物（miR－21－inhibitor）和阴性对照miRNA（negative control miRNA， NC－miRNA）转染入RSC96细胞中，构建出过表达miR－21的雪旺细胞株（ MI－SC）、抑制miR－21表达的雪旺细胞株（ IN－SC）及表达对照miRNA的雪旺细胞株（ NC－SC）；采用流式细胞仪检测3组细胞的凋亡情况； real－time PCR检测3组细胞中miR－21以及PTEN的表达情况；Western blot检测3组细胞中PTEN及凋亡相关蛋白激活型caspase－3（ cleaved caspase－3）的表达情况。结果：神经损伤组miR－21的表达量与对照组相比明显升高，神经损伤组PTEN mRNA的表达水平与对照组相比明显降低；与NC－SC相比，上调miR－21组细胞的凋亡比例减少，PTEN mRNA及蛋白的表达水平降低，cleaved caspase－3的表达水平降低，下调miR－21组细胞的凋亡比例增加，PTEN mRNA及蛋白的表达水平升高，cleaved caspase－3的表达水平升高（P＜0．05）。结论： miR－21可能通过下调PTEN的表达抑制雪旺细胞的凋亡，从而可能在周围神经的损伤修复中发挥作用。
目的：探討週圍神經損傷時，微小RNA－21（microRNA－21，miR－21）與雪旺細胞（SC）凋亡的關繫及相關分子機製。方法：採用實時熒光定量PCR（ real－time PCR）檢測動物模型中miR－21以及人第10號染色體缺失的燐痠酶及張力蛋白同源基因（phosphatase and tensin homologue deleted on chromosome 10，PTEN）的錶達情況；通過將miR－21類似物（miR－21－mimic）、miR－21抑製物（miR－21－inhibitor）和陰性對照miRNA（negative control miRNA， NC－miRNA）轉染入RSC96細胞中，構建齣過錶達miR－21的雪旺細胞株（ MI－SC）、抑製miR－21錶達的雪旺細胞株（ IN－SC）及錶達對照miRNA的雪旺細胞株（ NC－SC）；採用流式細胞儀檢測3組細胞的凋亡情況； real－time PCR檢測3組細胞中miR－21以及PTEN的錶達情況；Western blot檢測3組細胞中PTEN及凋亡相關蛋白激活型caspase－3（ cleaved caspase－3）的錶達情況。結果：神經損傷組miR－21的錶達量與對照組相比明顯升高，神經損傷組PTEN mRNA的錶達水平與對照組相比明顯降低；與NC－SC相比，上調miR－21組細胞的凋亡比例減少，PTEN mRNA及蛋白的錶達水平降低，cleaved caspase－3的錶達水平降低，下調miR－21組細胞的凋亡比例增加，PTEN mRNA及蛋白的錶達水平升高，cleaved caspase－3的錶達水平升高（P＜0．05）。結論： miR－21可能通過下調PTEN的錶達抑製雪旺細胞的凋亡，從而可能在週圍神經的損傷脩複中髮揮作用。
목적：탐토주위신경손상시，미소RNA－21（microRNA－21，miR－21）여설왕세포（SC）조망적관계급상관분자궤제。방법：채용실시형광정량PCR（ real－time PCR）검측동물모형중miR－21이급인제10호염색체결실적린산매급장력단백동원기인（phosphatase and tensin homologue deleted on chromosome 10，PTEN）적표체정황；통과장miR－21유사물（miR－21－mimic）、miR－21억제물（miR－21－inhibitor）화음성대조miRNA（negative control miRNA， NC－miRNA）전염입RSC96세포중，구건출과표체miR－21적설왕세포주（ MI－SC）、억제miR－21표체적설왕세포주（ IN－SC）급표체대조miRNA적설왕세포주（ NC－SC）；채용류식세포의검측3조세포적조망정황； real－time PCR검측3조세포중miR－21이급PTEN적표체정황；Western blot검측3조세포중PTEN급조망상관단백격활형caspase－3（ cleaved caspase－3）적표체정황。결과：신경손상조miR－21적표체량여대조조상비명현승고，신경손상조PTEN mRNA적표체수평여대조조상비명현강저；여NC－SC상비，상조miR－21조세포적조망비례감소，PTEN mRNA급단백적표체수평강저，cleaved caspase－3적표체수평강저，하조miR－21조세포적조망비례증가，PTEN mRNA급단백적표체수평승고，cleaved caspase－3적표체수평승고（P＜0．05）。결론： miR－21가능통과하조PTEN적표체억제설왕세포적조망，종이가능재주위신경적손상수복중발휘작용。
AIM:To explore the relationship and molecular mechanism between microRNA-21(miR-21) and Schwann cells ( SC) following peripheral nerve injury.METHODS: The mRNA expression of miR-21 and phosphatase and tensin homologue deleted on chromosome ten ( PTEN) in animal model were detected by real-time PCR.The over-ex-pression of miR-21 and inhibition of miR-21 expression in the Schwann cells according to transfection of lentiviral vectors were performed, the nonspecific miRNA was used as a negative control ( NC) .The cell apoptosis was measured by flow cy-tometry.The mRNA expression of miR-21 and PTEN in the cells was detected by real-time PCR.The protein expression of PTEN and cleaved caspase-3 was determined by Western blot.RESULTS: The level of miR-21 was significantly higher and the mRNA level of PTEN was significantly lower in the model of nerve injury than those in control group.miR-21 over-expression decreased the number of apoptotic Schwann cells compared with NC-SC.The mRNA expression of PTEN was down-regulated by over-expression of miR-21.The protein expression of PTEN and cleaved caspase-3 was down-regulated by over-expression of miR-21 (P<0.05).CONCLUSION: miR-21 may play an important role in the peripheral nerve injury through inhibiting apoptosis of Schwann cells by down-regulating the expression of PTEN.
